Analysis of machine perfusion benefits in kidney grafts: A preclinical study

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Abstract

Background: Machine perfusion (MP) has potential benefits for marginal organs such as from deceased from cardiac death donors (DCD). However, there is still no consensus on MP benefits. We aimed to determine machine perfusion benefits on kidney grafts.Methods: We evaluated kidney grafts preserved in ViaspanUW or KPS solutions either by CS or MP, in a DCD pig model (60 min warm ischemia + 24 h hypothermic preservation). Endpoints were: function recovery, quality of function during follow up (3 month), inflammation, fibrosis, animal survival.Results: ViaspanUW-CS animals did not recover function, while in other groups early follow up showed similar values for kidney function. Alanine peptidase and β-NAG activities in the urine were higher in CS than in MP groups. Oxydative stress was lower in KPS-MP animals. Histology was improved by MP over CS. Survival was 0% in ViaspanUW-CS and 60% in other groups. Chronic inflammation, epithelial-to-mesenchymal transition and fibrosis were lowest in KPS-MP, followed by KPS-CS and ViaspanUW-MP.Conclusions: With ViaspanUW, effects of MP are obvious as only MP kidney recovered function and allowed survival. With KPS, the benefits of MP over CS are not directly obvious in the early follow up period and only histological analysis, urinary tubular enzymes and red/ox status was discriminating. Chronic follow-up was more conclusive, with a clear superiority of MP over CS, independently of the solution used. KPS was proven superior to ViaspanUW in each preservation method in terms of function and outcome. In our pre-clinical animal model of DCD transplantation, MP offers critical benefits. © 2011 Vaziri et al; licensee BioMed Central Ltd.

Figures

  • Table 1 Solutions Composition
  • Figure 1 Perfusion Parameters and Kidney function following reperfusion. A: Flow rate and B: Resistance of machine perfused kidneys. C: Diuresis before and after transplantation. D: Serum creatinine before and after transplantation. E: Sodium excretion fraction. F: Glycosuria. G: Osmolarity ratio between blood and urine. Shown are mean ± SEM, statistics: † : p < 0.05 to ViaspanUW CS; * : p < 0.05 to ViaspanUW MP; ° : p < 0.05 to KPS CS; ¶ : p < 0.05 to KPS-MP.
  • Figure 2 Tubular integrity and Red/Ox Status following reperfusion. A: Alanine aminopeptidase activity in urine. B: b-Nacetylglucosaminidase activity in urine. C: Blood reduced gutathion over total glutathion ratio. Shown are mean ± SEM, statistics: † : p < 0.05 to ViaspanUW CS; * : p < 0.05 to ViaspanUW MP; ° : p < 0.05 to KPS CS; ¶ : p < 0.05 to KPS-MP.
  • Figure 3 Graft Histology. Representative PAS staining of kidney biopsies at day 7 and Month 1 post transplantation. LBB: loss of brush border; CD: Endoluminal cell detachment; Ti: tubulo-interstitial inflammation.
  • Table 2 Histological Evaluation
  • Figure 4 Inflammation. A: Representative images of typical ED1+ (top) and CD3+ (bottom) staining of kidneys from each group. B: graphical representation of the number of ED1 positive cells at each time point for each group. C: graphical representation of the number of CD3 positive cells at each time. Shown are mean ± SEM, statistics: † : p < 0.05 to ViaspanUW CS; * : p < 0.05 to ViaspanUW MP; ° : p < 0.05 to KPS CS; ¶ : p < 0.05 to KPS-MP.
  • Figure 5 EMT development. A, B, C and D: Representative staining for Vimentin at 3 months. E: quantification of staining in each group. Shown are mean ± SEM, statistics: † : p < 0.05 to ViaspanUW CS; * : p < 0.05 to ViaspanUW MP; ° : p < 0.05 to KPS CS; ¶ : p < 0.05 to KPS-MP.
  • Figure 6 3 month Outcome. Survival was measured and represented by a Kaplan-Meier plot (A). Function was determined: Creatinemia (B) and proteinuria (C) Representative images of Sirius Red staining of sections obtained from Control (D), ViaspanUW-MP (E), KPS-CS (F) KPS-MP (G) kidneys. Original magnification x100. H: Quantification of fibrosis in kidneys from each group studied. Shown are mean ± SEM, statistics: † : p < 0.05 to ViaspanUW CS; * : p < 0.05 to ViaspanUW MP; ° : p < 0.05 to KPS CS; ¶ : p < 0.05 to KPS-MP.

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Vaziri, N., Thuillier, R., Favreau, F. D., Michel Eugene, Milin, S., Chatauret, N. P., … Barrou, B. (2011). Analysis of machine perfusion benefits in kidney grafts: A preclinical study. Journal of Translational Medicine, 9. https://doi.org/10.1186/1479-5876-9-15

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